五味子乙素对脂多糖诱导的心肌H9c2细胞炎性损伤的影响及其作用机制研究

Impact of Schisandrin B on LPS-induced Inflammatory Injury of Myocardial H9c2 Cells and the Action Mechanism

目的探讨五味子乙素对脂多糖(LPS)诱导的心肌H9c2细胞炎性损伤的影响及其作用机制,以期为五味子乙素的临床应用奠定基础。方法 2017年3月—2018年1月,将传代培养的心肌H9c2细胞接种于96孔板上并分为正常对照组(A组)、LPS诱导组(B组)、LPS+低剂量五味子乙素组(C组)、LPS+中剂量五味子乙素组(D组)、LPS+高剂量五味子乙素组(E组),细胞生长贴壁后A组心肌H9c2细胞常规培养,B组心肌H9c2细胞仅加入含1 mg/L的LPS,C、D、E组心肌H9c2细胞在B组基础上分别加入10、50、150 mg/L五味子乙素,5组心肌H9c2细胞均培养24 h。比较5组心肌H9c2细胞相对存活率、凋亡率、炎性因子含量及PI3K、Akt、p-Akt、FoxO1、p-FoxO1蛋白相对表达量。结果 (1)C、D、E组心肌H9c2细胞相对存活率高于B组,D、E组心肌H9c2细胞相对存活率高于C组,E组心肌H9c2细胞相对存活率高于D组(P<0.05)。(2)B、C、D、E组心肌H9c2细胞凋亡率高于A组,C、D、E组心肌H9c2细胞凋亡率低于B组,D、E组心肌H9c2细胞凋亡率低于C组,E组心肌H9c2细胞凋亡率低于D组(P<0.05)。(3)B、C、D、E组心肌H9c2细胞白介素6(IL-6)、肿瘤坏死因子α(TNF-α)含量高于A组,C、D、E心肌H9c2细胞IL-6、TNF-α含量低于B组,D、E组心肌H9c2细胞IL-6、TNF-α含量低于C组,E组心肌H9c2细胞IL-6、TNF-α含量低于D组(P<0.05)。(4)5组心肌H9c2细胞Akt、FoxO1蛋白相对表达量比较,差异无统计学意义(P>0.05);B、C、D组心肌H9c2细胞PI3K、p-Akt、p-FoxO1蛋白相对表达量低于A组,C、D、E组心肌H9c2细胞PI3K、p-Akt、p-FoxO1蛋白相对表达量高于B组,D、E组心肌H9c2细胞PI3K、p-Akt、p-FoxO1蛋白相对表达量高于C组,E组心肌H9c2细胞PI3K、p-Akt、p-FoxO1蛋白相对表达量高于D组(P<0.05)。结论五味子乙素可有效促进LPS诱导的心肌H9c2细胞增殖、抑制心肌H9c2细胞凋亡、降低心肌H9c2细胞炎性因子含量,有利于减轻细胞炎性损伤,其作用机制可能与激活PI3K/Akt/FoxO1信号通路有关。

Objective To investigate the impact of Schisandrin B on LPS-induced inflammatory injury of myocardial H9c2 cells and the action mechanism.Methods From March 2017 to January 2018,serial sub-cultivated H9c2 cells were inoculated on 96-well plates and divided into A group,B group,C group,D group and E group,thereinto cells in A group received routine culture,cells in B group received LPS(1 mg/L),cells in C group,D group and E group received Schisandrin B with different concentration(10,50 and 150 mg/L,respectively)based on that of B group,cells in the four group cultivated for 24 hours.Relative survival rate,apoptosis rate,inflammatory cytokines contents,relative protein expression quantity of PI3K,Akt,p-Akt,FoxO1 and p-FoxO1 were compared in the five groups.Results (1)Relative survival rate in C group,D group and E group was statistically significantly higher than that in B group,respectively,relative survival rate in D group and E group was statistically significantly higher than that in C group,respectively,meanwhile relative survival rate in E group was statistically significantly higher than that in D group(P<0.05).(2)Apoptosis rate in B group,C group,D group and E group was statistically significantly higher than that in A group,respectively,apoptosis rate in C group,D group and E group was statistically significantly lower than that in B group,respectively,apoptosis rate in D group and E group was statistically significantly lower than that in C group,respectively,meanwhile apoptosis rate in E group was statistically significantly lower than that in D group(P<0.05).(3)IL-6 and TNF-αcontents in B group,C group,D group and E group were statistically significantly higher than those in A group,IL-6 and TNF-αcontents in C group,D group and E group were statistically significantly lower than those in B group,IL-6 and TNF-αcontents in D group and E group were statistically significantly lower than those in C group,meanwhile IL-6 and TNF-αcontents in E group were statistically significantly lower than those in D group(P<0.05).(4)No statistically significant difference of relative protein expression quantity of Akt or FoxO1 was found in the five groups(P>0.05);relative protein expression quantity of PI3K,p-Akt and p-FoxO1 in B group,C group and D group was statistically significantly lower than that in A group,respectively,relative protein expression quantity of PI3K,p-Akt and p-FoxO1 in C group,D group and E group was statistically significantly higher than that in B group,respectively,relative protein expression quantity of PI3K,p-Akt and p-FoxO1 in D group and E group was statistically significantly higher than that in C group,respectively,meanwhile relative protein expression quantity of PI3K,p-Akt and p-FoxO1 in E group was statistically significantly higher than that in D group,respectively(P<0.05).Conclusion Schisandrin B can effectively promote the proliferation,inhibit the apoptosis and reduce the inflammatory cytokines content of LPS-induced myocardial H9c2 cells,is helpful to relieve the inflammatory injury,its action mechanism may correlated with activation of PI3K/Akt/FoxO1 signal pathway.